Observatorio de I+D+i UPM

Memorias de investigación
Research Publications in journals:
Novel lambda FRET spectral confocal microscopy imaging method
Year:2009
Research Areas
  • Electronics engineering
Information
Abstract
We report a highly specific, sensitive, and robust method for analyzing fluorescence resonance energy transfer (FRET) based on spectral laser scanning confocal microscopy imaging. The lambda FRET (kFRET) algorithm comprises imaging of a FRET sample at multiple emission wavelengths rendering a FRET spectrum, which is separated into its donor and acceptor components to obtain a pixel-based calculation of FRET efficiency. The method uses a novel off-line precalibration procedure for spectral bleed-through correction based on the acquisition of reference reflection images, which simplifies the method and reduces variability. kFRET method was validated using structurally characterized FRET standards with variable linker lengths and stoichiometries designed for this purpose. kFRET performed better than other well-established methods, such as acceptor photobleaching and sensitized emission-based methods, in terms of specificity, reproducibility, and sensitivity to distance variations. Moreover, kFRET analysis was unaffected by high fluorochrome spectral overlap and cellular autofluorescence. The kFRET method demonstrated outstanding performance in intra- and intermolecular FRET analysis in both fixed and live cell imaging studies.
International
Si
JCR
Si
Title
MICROSCOPY RESEARCH AND TECHNIQUE
ISBN
1059-910X
Impact factor JCR
1,883
Impact info
Volume
72
10.1002/jemt.20633
Journal number
1
From page
1
To page
11
Month
ENERO
Ranking
Participants
  • Autor: Andres de Santos Lleo (UPM)
Research Group, Departaments and Institutes related
  • Creador: Grupo de Investigación: Tecnología de imágenes biomédicas
  • Centro o Instituto I+D+i: Centro de tecnología Biomédica CTB
  • Departamento: Ingeniería Electrónica
S2i 2019 Observatorio de investigación @ UPM con la colaboración del Consejo Social UPM
Cofinanciación del MINECO en el marco del Programa INNCIDE 2011 (OTR-2011-0236)
Cofinanciación del MINECO en el marco del Programa INNPACTO (IPT-020000-2010-22)